Sequence-specific assignments in NMR spectra of paramagnetic systems: A non-systematic approach

PRE‐driven protein NMR structures: an alternative approach in highly paramagnetic systems

“…The complete resonance assignment of the protein was obtained combining the conventional approach based on triple resonance experiments (Table S1) with a non-systematic procedure using a combination of 1D NOEs, 13 C direct detection, double and triple resonance experiments recorded with parameters optimized a-la-carte (Table S2) [92]. These experiments provided the complete NMR assignment of PioC (BRMB entry 34487) [93].…”

“…The set of NMR experiments used for sequence specific assignment, NOE collection, and 15 N relaxation analysis is summarized in TableS1. To identify signals affected by the hyperfine interaction, tailored experiments were performed [64,91,92]. Experimental parameters are summarized in TableS2.…”

Paramagnetic NMR Spectroscopy Is a Tool to Address Reactivity, Structure, and Protein–Protein Interactions of Metalloproteins: The Case of Iron–Sulfur Proteins

“…This optimization strategy can be easily applied to all available triple-resonance experiments, allowing one to record "paramagnetic" versions of, virtually, any triple-resonance experiment. As shown in Figure 3B, an HNCA optimized as above permits the identification of the HNCA peaks of three out of the four cluster-bound cysteine residues that, by no means, are observable using a routine HNCA setup [86]. [87].…”

Biochemical and cellular characterization of the CISD3 protein: Molecular bases of cluster release and destabilizing effects of nitric oxide

“…Among the other residues showing chemical shift perturbation above the threshold level, Phe 70 (N-terminal domain) and the corresponding Tyr 108 (C-terminal domain) showed a very similar shift difference (0.092 versus 0.088). The paramagnetism of the cluster impedes the identification of NMR signals of residues close to the [Fe 2 S 2 ] clusters, even when tailored approaches are used ( 44 , 45 ), as a consequence we do not have enough data for a reliable comparison among the two cluster sites. However, Figure 6 B shows that larger perturbations occur in the C-terminal CDGSH domain and, therefore, the two cluster sites might not be equivalent.…”

“…To provide atomic-level insights into the proximity of the two clusters, we performed 1 H NMR spectra of the reduced and oxidized CISD3. Paramagnetically shifted 1 H NMR signals constitute a very unique and sensitive fingerprint for the characterization of the first coordination sphere ( 44 , 45 , 46 , 47 ). As shown in Figure 7 , oxidized CISD3 shows five hyperfine shifted signals spread between ca.…”